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Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
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Quantification of Metabolic Products from Microbial Hosts in Complex Media Using Optically Diffracting Hydrogels.

Sukwon Jung1, Kelsey I MacConaghy1, Michael T Guarnieri2

  • 1Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado 80303, United States.

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We developed photonic crystal hydrogels for real-time screening of microbial biocatalysts. This method enables high-throughput detection of bioproducts like alcohols and organic acids in biological samples.

Keywords:
crystalline colloidal arrayethanolgenome engineeringmetabolic engineeringorganic acidsphotonic crystalsynthetic biology

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Area of Science:

  • Biotechnology and Bioengineering
  • Materials Science
  • Chemical Engineering

Background:

  • Screening microbial biocatalysts for enhanced bioproducts is crucial for industrial biotechnology.
  • Conventional methods for detecting bioproducts often lack real-time capabilities or sensitivity in complex media.
  • Developing novel sensing platforms is essential to overcome these limitations.

Purpose of the Study:

  • To present a versatile platform for in situ and real-time screening of microbial biocatalysts using photonic crystal hydrogels.
  • To demonstrate the utility of this approach for detecting alcohols and organic acids.
  • To enable high-throughput screening of bioproducts in complex biological media.

Main Methods:

  • Fabrication of optically diffracting hydrogel films based on polymerized N-isopropylacrylamide in a microwell plate format.
  • Optical characterization of hydrogel films using a microwell plate reader for high-throughput screening.
  • Correlation of hydrogel response with the octanol-water partition coefficient (log P) of analytes.
  • Detection and quantification of ethanol and succinic acid secreted by microbial strains.

Main Results:

  • The hydrogel films exhibited optical responses to a broad range of alcohols and organic acids.
  • Hydrogel film response strongly correlated with the analyte's octanol-water partition coefficient (log P).
  • Differential secretion of ethanol and succinic acid by Zymomonas mobilis and Actinobacillus succinogenes, respectively, was detected with high sensitivity (down to ~3 g/L).
  • Results were validated using high-performance liquid chromatography.

Conclusions:

  • The photonic crystal hydrogel platform offers a versatile and sensitive method for real-time bioproduct detection.
  • This approach facilitates high-throughput screening of microbial biocatalysts in complex biological samples.
  • The technology overcomes challenges associated with conventional optical sensing methods for small organic analytes.